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Cancer immunotherapy based on image-guided STING activation by nucleotide nanocomplex-decorated ultrasound microbubbles

Abstract

The cytosolic innate immune sensor cyclic GMP-AMP synthase-stimulator of interferon genes (cGAS-STING) pathway is crucial for priming adaptive antitumour immunity through antigen-presenting cells (APCs). Natural agonists, such as cyclic dinucleotides (CDNs), activate the cGAS-STING pathway, but their clinical translation is impeded by poor cytosolic entry and serum stability, low specificity and rapid tissue clearance. Here we developed an ultrasound (US)-guided cancer immunotherapy platform using nanocomplexes composed of 2′3′-cyclic guanosine monophosphate-adenosine monophosphate (cGAMP) electrostatically bound to biocompatible branched cationic biopolymers that are conjugated onto APC-targeting microbubbles (MBs). The nanocomplex-conjugated MBs engaged with APCs and efficiently delivered cGAMP into the cytosol via sonoporation, resulting in activation of cGAS-STING and downstream proinflammatory pathways that efficiently prime antigen-specific T cells. This bridging of innate and adaptive immunity inhibited tumour growth in both localized and metastatic murine cancer models. Our findings demonstrate that targeted local activation of STING in APCs under spatiotemporal US stimulation results in systemic antitumour immunity and improves the therapeutic efficacy of checkpoint blockade, thus paving the way towards novel image-guided strategies for targeted immunotherapy of cancer.

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Fig. 1: Nanocomplex-decorated microbubbles targeting CD11b on APCs.
Fig. 2: Sonoporation of APCs targeted by ncMBs (MUSIC) induces activation of STING-IRF signalling in vitro.
Fig. 3: MUSIC activates STING signalling and T cell response in primary breast cancer in vivo.
Fig. 4: MUSIC activates STING-mediated antitumour immunity.
Fig. 5: MUSIC activates systemic antitumour immunity to inhibit breast cancer metastasis.

Data availability

The authors declare that data supporting the findings of this study are available within the article and its Supplementary Information. The datasets generated and analysed during the study are publicly available at https://osf.io/phcmx. Source data are provided with this paper.

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Acknowledgements

This work was supported in part by the Cancer Prevention and Research Institute of Texas (CPRIT) grants RR150010, RP210199 and RP19023, the Department of Defense grants W81XWH-21-1-0332/0333 and W81XWH-17-1-0401, the Susan G. Komen Foundation Career Catalyst Research grant CCR19605871 and the National Cancer Institute grant 1K08CA241070. R.F.M. is a CPRIT Established Investigator. Research reported in this publication was also supported by the Children’s Cancer Fund Comprehensive Center for Pediatric Oncology Research. The authors acknowledge Siemens Medical Solutions USA for the Siemens Sequoia ultrasound scanner loan. The authors acknowledge the UT Southwestern Harold C. Simmons Cancer Center Support grant P30 CA142543 for the support provided by the Small Animal Imaging shared resource. The authors also would like to acknowledge Erin Moore (Creatives Services, Department of Radiology, The University of Texas Southwestern Medical Center) for the illustrations in Fig. 1a,b and Supplementary Fig. 5a, and C. Wogan from the MD Anderson Cancer Center for editorial help.

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Authors and Affiliations

Authors

Contributions

W.J. and J.L. conceived the project and were responsible for all phases of the research. X.L., S.K., Y.W., W.J. and J.L. designed the experiments. S.K., C.d.G.L., R.F.M. and J.L. conceived the microbubble platform. R.F.M. and J.L. provided guidance for ultrasound experiments. S.K. and N.N. prepared and characterized the MUSIC platform. X.L. and S.K. performed the in vitro experiments. X.L., S.K., M.Y. and J.S. performed the in vivo experiments. X.L. and S.K. collected the data. X.L., S.K., Y.W., K.H., D.L., Y.L., R.G., B.Y.S.K., W.J. and J.L. analysed and interpreted the data. X.L., S.K., Y.W., W.J. and J.L. performed the literature review. The manuscript was written through contributions of all authors. All authors have given approval to the final version of the manuscript.

Corresponding authors

Correspondence to Wen Jiang or Jacques Lux.

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Competing interests

A provisional patent application based on the technology described in the manuscript has been filed by The University of Texas Southwestern Medical Center, with S.K., J.L. and W.J. as inventors, application number 63/173,956. All other authors declare no competing interests.

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Nature Nanotechnology thanks Christopher Jewell and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–30 and Table 1.

Reporting Summary

Supplementary Video 1

cGAMP-loaded SpeDex MBs-aCD11b (ncMBs) were visualized using US during intratumoural injection.

Source data

Source Data Fig. 2

Unprocessed western blots.

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Li, X., Khorsandi, S., Wang, Y. et al. Cancer immunotherapy based on image-guided STING activation by nucleotide nanocomplex-decorated ultrasound microbubbles. Nat. Nanotechnol. (2022). https://doi.org/10.1038/s41565-022-01134-z

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